Proteomic study of hippocampal tissue on electroacupuncture‑improved learning and memory impairment in Alzheimer's disease rats based on tandem mass tag labeling

KE Chao; SHAN Shengtao; CAO Yang; TAN Yan; XIE Zhengrong; ZHOU Zhuo; YUAN Liumei; PAN Jiang; ZHANG Wei

doi:10.16305/j.1007-1334.2026.z20250323001

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Proteomic study of hippocampal tissue on electroacupuncture‑improved learning and memory impairment in Alzheimer's disease rats based on tandem mass tag labeling

更新时间：2026-01-06

- Proteomic study of hippocampal tissue on electroacupuncture‑improved learning and memory impairment in Alzheimer's disease rats based on tandem mass tag labeling
- Shanghai Journal of Traditional Chinese Medicine Vol. 60, Issue 1, Pages: 12-20(2026)
- 作者机构：
  
  湖南中医药大学第一附属医院（湖南长沙 410007）
- 作者简介：
- 基金信息：
- DOI：10.16305/j.1007-1334.2026.z20250323001
  CLC：
- Received：23 March 2025，
  
  Published：10 January 2026
- 稿件说明：
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柯超,单生涛,曹洋,等.基于串联质谱标签的电针改善阿尔茨海默病大鼠学习记忆损伤的海马组织蛋白质组学研究[J].上海中医药杂志,2026,60(1):12-20.

KE Chao,SHAN Shengtao,CAO Yang,et al.Proteomic study of hippocampal tissue on electroacupuncture‑improved learning and memory impairment in Alzheimer's disease rats based on tandem mass tag labeling[J].Shanghai Journal of Traditional Chinese Medicine,2026,60(1):12-20.
柯超,单生涛,曹洋,等.基于串联质谱标签的电针改善阿尔茨海默病大鼠学习记忆损伤的海马组织蛋白质组学研究[J].上海中医药杂志,2026,60(1):12-20. DOI： 10.16305/j.1007-1334.2026.z20250323001.

KE Chao,SHAN Shengtao,CAO Yang,et al.Proteomic study of hippocampal tissue on electroacupuncture‑improved learning and memory impairment in Alzheimer's disease rats based on tandem mass tag labeling[J].Shanghai Journal of Traditional Chinese Medicine,2026,60(1):12-20. DOI： 10.16305/j.1007-1334.2026.z20250323001.

摘要

目的

观察电针对阿尔茨海默病（AD）大鼠学习记忆能力和海马组织蛋白质组的影响，探索电针防治AD的差异蛋白及其调控途径。

方法

将24只SD大鼠按照随机数字法分为假手术（Sham）组、模型（Model）组、电针（EA）组，每组8只。采用双侧大脑海马CA1区内注射淀粉样蛋白β蛋白片段1-42（Aβ1-42）构建AD模型。连续电针干预7周后，采用Morris水迷宫实验评价大鼠学习记忆能力，采用新物体识别实验评估大鼠识别记忆和新奇偏好能力，采用串联质谱标签（TMT）标记定量蛋白质组学技术检测大鼠海马组织蛋白质组，对差异蛋白进行基因本体（GO）和京都基因与基因组数据库（KEGG）等富集分析，采用平行反应监测（PRM）法对关键差异蛋白进行验证。

结果

与Sham组比较，Model组大鼠的逃避潜伏期显著延长（

0.01），穿越平台次数显著减少（

0.01），目标象限停留时间均显著缩短（

0.01）；与Model组相比，EA组的逃避潜伏期明显短于Model组（

0.01），EA组穿越平台次数显著增加（

0.01），目标象限停留时间则显著延长（

0.01）。各组大鼠游泳速度没有显著的组间差异（

0.05）。在相同物体探索阶段，各组大鼠探索相同物体识别指数没有显著差异（

0.05）；在新物体探索阶段，与Sham组比，Model组大鼠新物体识别指数明显下降（

0.01），与Model组相比，EA组新物体识别指数明显升高（

0.01）。蛋白质组学分析结果显示，共鉴定出211种差异表达蛋白，根据差异筛选出11个显著逆转的目的蛋白；KEGG分析结果表明，突触囊泡周期、谷氨酸能突触通路为主要调控通路。最终确定了两种关键蛋白，即蛋白酪氨酸磷酸酶非受体5型（PTPN5）及钠和氯依赖的γ-氨基丁酸转运体3（GAT3），它们分别与谷氨酸能突触通路和突触囊泡通路相关。PRM结果表明，与Sham组相比，Model组大鼠海马组织中PTPN5、GAT3蛋白的表达量增加（

0.01）；与Model组相比，EA组大鼠海马组织中PTPN5、GAT3蛋白表达量减少（

0.01），与蛋白质组学趋势一致。

结论

电针可能通过调控海马组织PTPN5、GAT3的表达，进而改善AD大鼠的学习记忆能力。

Abstract

Objective

To investigate the effects of electroacupuncture （EA） on learning and memory abilities and hippocampal tissue proteome in Alzheimer's disease （AD） rats， and to explore differential proteins and regulatory pathways involved in the prevention and treatment of AD by EA.

Methods

Twenty-four SD rats were randomly divided into sham operation （Sham） group， Model group， and EA group， with 8 rats in each group. The AD model was established through injection of amyloid β protein 1-42 into bilateral hippocampal CA1 region. Following 7-week EA intervention， the learning and memory abilities of rats were evaluated using the Morris water maze experiment， and the recognition memory and novelty preference abilities of rats were assessed through the novel object recognition experiment. Hippocampal proteomic profiling was performed through tandem mass tag-labeling quantitative proteomics. Differential proteins underwent Gene Ontology（GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analyses. The key differential proteins were verified using the parallel reaction monitoring （PRM） method.

Results

Compared with the Sham group， the rats in the Model group exhibited significantly prolonged escape latency （

0.01）， reduced platform crossings numbers（

0.01）， and decreased target quadrant dwell time （

0.01）. Compared with the Model group， the escape latency of rats in the EA group was significantly shorter than that of the Model group （

0.01）， the number of platform crossings was significantly increased （

0.01）， and the target quadrant dwell time was significantly prolonged （

0.01）. There was no significant difference in swimming speed among the different groups of rats （

0.05）. During the exploration of the same objects， there was no significant difference in the recognition index of the same objects among all groups of rats （

0.05）； during the exploration of new objects， compared with the Sham group， the recognition index of new objects in the Model group was significantly decreased （

0.01）， and compared with the Model group， the recognition index of novel objects in the EA group was significantly increased （

0.01）. The proteomic analysis results showed that a total of 211 differentially expressed proteins were identified. Based on these differences， 11 significant reversal target proteins were selected. KEGG enrichment results highlighted synaptic vesicle cycle and glutamatergic synapse pathways as primary regulatory mechanisms. Two key proteins were finally identified， namely protein tyrosine phosphatase non-receptor type 5 （PTPN5） as well as sodium and chloride-dependent γ-aminobutyric acid transporter 3 （GAT3）， which were associated with the glutamatergic synaptic pathway and the synaptic vesicle path

way respectively. The PRM results indicated that compared with the Sham group， the expressions of PTPN5 and GAT3 proteins in the hippocampal tissue of the Model group were increased （

0.01）； compared with the Model group， the expressions of PTPN5 and GAT3 proteins in the hippocampal tissue of the EA group were decreased （

0.01）， which was consistent with the proteinomics trends.

Conclusions

EA may improve the learning and memory abilities of AD rats by regulating the expressions of PTPN5 and GAT3 in the hippocampal tissue.

关键词

Keywords

references

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